1058 DIFFERENCES IN DNA METHYLATION PATTERN OF PRIMARY BLADDER TUMOURS CORRELATE WITH THEIR METASTATIC POTENTIAL

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You have accessJournal of UrologyBladder Cancer: Basic Research III1 Apr 20121058 DIFFERENCES IN DNA METHYLATION PATTERN OF PRIMARY BLADDER TUMOURS CORRELATE WITH THEIR METASTATIC POTENTIAL Beatrice Stubendorff, Eva Dudziec, James Catto, Jimsgene Sanjmyatav, Mieczeslaw Gajda, Heiko Wunderlich, Marc-Oliver Grimm, and Kerstin Junker Beatrice StubendorffBeatrice Stubendorff Jena, Germany More articles by this author , Eva DudziecEva Dudziec Sheffiled, United Kingdom More articles by this author , James CattoJames Catto Sheffiled, United Kingdom More articles by this author , Jimsgene SanjmyatavJimsgene Sanjmyatav Jena, Germany More articles by this author , Mieczeslaw GajdaMieczeslaw Gajda Jena, Germany More articles by this author , Heiko WunderlichHeiko Wunderlich Jena, Germany More articles by this author , Marc-Oliver GrimmMarc-Oliver Grimm Jena, Germany More articles by this author , and Kerstin JunkerKerstin Junker Jena, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1164AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The prognosis of patients with metastatic bladder cancer is poor with a 5-year survival rate of only 6%. Early prediction of the metastatic risk in primary tumors can lead to improvements in prognosis and therapy. Currently, there are no parameters available that enable an individual risk assessment for patients with metastatic bladder cancer. Therefore, identification of new reliable prognostic markers for early prediction of tumor spread is required. The aim of this project is to determine whether changes in DNA methylation correlate with the metastasis risk and to identify a specific DNA methylation pattern that provides a reliable tool for prognosis assessment. METHODS Genomic DNA was isolated from 23 invasive bladder tumour tissues with and without lymph node metastases. For enrichment of methylated fragments genomic DNA was incubated with 5-Methylcytosin antibody. Input and IP were labelled with Cy3 and Cy5. Labelled DNA was hybridizes onto CpG island microarrays. Cluster analysis enabled selection of differentially methylated candidate genes. The actual methylation state was confirmed by cleavage of genomic DNA with methylation dependent restriction enzyme (McrBc) followed by quantitative PCR. RESULTS In total, 201 promoter CpG islands show highly significant differences in DNA methylation between metastasised and non-metastasised primary bladder tumors (p<0,01). In metastasised tumors 74 CpG islands are hypermethylated and 127 CpG islands are hypomethylated. Data base research for identification of candidate genes resulted in 61 hypermethylated and 58 hypomethylated genes that can clearly assigned to the identified promoter associated CpG islands. Validation of methylation level of candidate gene RASSF1 resulted in higher methylation in metastatic tumors (mean methylation: 38%) than in non-metastatic tumors (mean methylation: 26%). RASSF1 methylation shows strong correlation to T stage (r=0,65). Methylation of candidate gene KISS1R shows significant differences between patient groups (p=0,001). Mean methylation in metastatic tumours is 48% and in non-metastatic tumours 22%. CONCLUSIONS Primary bladder tumors show significant differences in their DNA methylation pattern in correlation to their metastatic potential. KISS1R methylation is strongly associated with metastatic risk of primary tumors. Further analysis will demonstrate the correlation between changes in DNA methyaltion and expression of corresponding genes. The impact of DNA methylation on tumor behaviour shall be determined in functional analysis. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e428-e429 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Beatrice Stubendorff Jena, Germany More articles by this author Eva Dudziec Sheffiled, United Kingdom More articles by this author James Catto Sheffiled, United Kingdom More articles by this author Jimsgene Sanjmyatav Jena, Germany More articles by this author Mieczeslaw Gajda Jena, Germany More articles by this author Heiko Wunderlich Jena, Germany More articles by this author Marc-Oliver Grimm Jena, Germany More articles by this author Kerstin Junker Jena, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...